Parabiotic chamber apparatus for cell culture studies



P- A. BRETT Dec. 7, 1965 PARABIOTIC CHAMBER APPARATUS FOR CELL CULTURESTUDIES Filed Sept. 21, 1962 INVENTOR. PERRY A. 8967'? ATTORNEY UnitcdStates Patent 3,221,878 PARABIOTIC CHAMBER APPARATUS FOR CELL CULTURESTUDIES lPerry A. Brett, Levittown, Pa., assignor to Bellco Glass,

Incorporated, Vineland, Ni, a corporation of New .l'ersey Filed Sept.21; 1962', Ser. No. 225,323 4- Claims. (Cl. 210-94).

The presentinventionis directed to parabiotic chamber apparatus for cellculture studies.

The use of conventional tissue culture glassware in the study of cellinterrelationships. has a distinct disadvantage when directcontact ofdifferent types of cells is undesirable. Thus, instudies of-morphologicchanges in neoplastic cells, it isfrequently essential that singlecelllayerson thin glassmounts be manipulated in growth vessels, but withoutdirect contactof the different types of cells that arebeing used. Forexample, this is necessitated in experiments in which freeexchangeofsubcellular sized particles is desired, but free exchange of cells is tobe avoided.

The present invention has as an object the provision of a parabioticchamber apparatus forcellculture-studies, and the like.

The present invention has as another object the provision of apparatusin which minute particles may be exchanged between vessels-without thepossibility-of the exchange of larger particles.

Other objects will appear hereinafter.

For the; purpose of illustrating the invention, there are shown in thedrawings forms whichare presently preferred; it being understood,however, that thisinvention is not limited to the precise arrangementsand instrumentalities shown.

Referringto the drawings, wherein like reference characters refer tolike parts:

FIGURE 1 is a perspective view-of one embodiment of the parabioticchamber apparatus for cell culture studiesof the present invention.

FIGURE 2 is a longitudinal sectional view takenon line 2-2 of FIGURE 1.

FIGURE 3 is a cross sectional view taken on line 33 of FIGURE 2.

FIGURE 4 is a fragmentary sectional View of another embodiment ofthe-parabiotic chamber apparatus .forcell culture studies of the presentinvention.

Referring initially to FIGURES I through 3 inclusive, there is disclosedtherein the parabiotic chamber apparatus designated generally as 10,which includes the holder 12, the growth vessels Hand 16-, with theirrespective plugs 13 and 2t and'the dialysis membrane 22.

The holder 12 includes a base 24- of stamped sheet metal or the like.The base 24 is provided with the clamping members 26 and 28, whichextendupwardly from the middle portion of the base 24;

The clamping member 26 is essentiallyan angle iron with its short arm 30welded'or otherwise metallurgically.

joined to the base 24: The upright arm 32Xof clamping.

member 26 isslotted at 34 to-providethe fingers 36and 38, The bightofthe slot 34' is offset to provide the retention shoulder 40; Theclamping member 26 is a.

relatively rigid and inflexible member.

The clamping member 28 may be formed of two elements, namely the springelement. 42, and-the gripperelement 44, with the elements 42 and 44being welded or otherwise metallurgically joined together. The springelement 42 is welded or otherwise metallurgically joined to the top ofthe base 24.

The spring element 42 spring-urges the gripper element 44 towards theupright arm 32 of the clamping member 26.

The gripper element 44is slottedat 46, with the bight portion of theslot 46 being offset to provide a retention shoulder 48. The slot 46dividesthe gripper element 44 into fingers 50 and 52;

The fingers 36 and 50, the slots 34' and 46, and the fingers 38 and 52:of the respective clamping members 26 and28jare in juxtaposed alignment.As heretofore noted, the spring element42 urges the gripper element 44-towards the upright arm 32' of the clamping member 26. Preferably, theentire holder 12 is formed of stainless steel. However, a variety ofmetals, plastics, and other materials may be used for the manufactureofthe holder 12.

The growth vessels 14 andj16, and their respective plugs 18 and 20, areidentical. Hence, although the description of the growth vessel whichfollows will be limitedto growth vessel 14, it willapply with equalforce to growthvessel 1 6.

The growth vessel 14 may be formed from an originally substantiallycylindrical glass member, and is open at each of its ends. The end 54adjacent the plug 18 is tapered. The plug 18 which mayv be formed fromsilicone, rubber, synthetic rubber or like elastomer, etc., is matinglytapered to the end 54 so that a gas-tight fit may be secured betweentheend 54 and the plug 18.

The opposite end 56 ,of the. growth vessel 14 is flared outwardly froma.radiallyinwardly, extending neck 58, and-is provided at its outermostendwith a radially outwardly extending flange 60. Preferably, the faceof the flange 60 whichoperatively engages the dialysis membrane 22,namely the face, most remote from the end 54, is ground.

A dam 62 comprising an inwardly extending wall having' a straightuppermost edge is provided in growth vessel 14at a spaced distance fromthe tapered end 54. The height of thev top of the dam 62 verticallymeasured from the plane ofthefloorof the fiat 64 when the growth vessel14. is in the position. shown in FIGURE 2, is approximately equal to thevertical height of the top of the neck 53, at that portion of the neck58 which is bisected by the vertical plane of symmetry of the growthvessel 14.

The. fiat 64 extends for an appreciable distance in the middle portionotthe, growth vessel 14. As shown in the drawings (which are drawn toscale for a specific embodiment of the present invention), and inparticular in FIGURE 3, the flat 64 has a fiat undersurface whichextends laterally foralmost the entire outside diametric width of thecentral tubular portion of the growth vessel 14. It will be seenfrom-FIGURE 2 that longitudinally the flat 64 is defined by the dam 62at its end which is proximate the tapered end-54,' andeby a rise 66which is proximate the neck 58. Alternatively, the rise 66 may beeliminated and the flat may extend to the neck 58, and hence the fiat 64:be defined longitudinally by the neck 58 and the;dam 62;

The dialysis membrane-22 may be formed of any suitable material-forthispurpose. A large number of; suitable materialsare-available, and theappropriate one will be selected by the researchinvestigator to fit hisparticular need. Suitable dialysis membranes include mem-. branesprepared from cellulose, collodion, Millipore' fil ter, etc.Suchmaterials will not permitthe exchange of a tissue cell,- but willpermit the exchange of 'subcellular. particles. In the case of colloidalsolutions;(and the apparatus of: the present invention is useful forresearch in connection with such materials) the dialysis membrane willnot permit the passage therethrough of the colloid.

The dialysis membrane 22 is maintained in its operative positionintermediate the ground end faces of the Patented Dec. 7', 1965- 3flanges 60 of the growth vessels 14 and 16. Preferably, a minor amountof silicone grease or the like is provided on the ground end faces ofthe flanges 60 to secure a gastight seal.

The operation of the parabiotic chamber apparatus of the presentinvention is as follows:

The apparatus is assembled after the ground end faces of the flanges 60of the growth vessels 14 and 16 have been lightly coated with siliconegrease. Thus, the. dialysis membrane 22 is inserted intermediate thegrowth vessels 14 and 16. The growth vessels 14 and 16 are held in axialalignment, with their flats 64 lying in the same plane, and parallel tothe plane of the web of the base 24. The alignment is maintained by thespring action of the clamping member 28 against the clamping member 26.The insertion of the growth vessel 14 within the slot 46 and theinsertion of the growth vessel 16 within the slot 34 are facilitated bythe otfsetting of the fingers 36, 38, 50 and 52, which aid in themanipulation of the clamping member 28.

The offset retention shoulders 40 and 48 engage the ends 56 of thegrowth vessels 14 and 16 in the manner most clearly shown in FIGURE 2.Thus, the juxtaposed neck portions of the flanges 60 of the growthvessels 14 and 16 are engaged with the retention shoulders 40 and 48.

When dealing with tissue cultures, the cells are preferably transferredto the flats 64 of the growth vessels 14 and 16 as single cell layers onthin glass mounts, such as cover glasses. These are inserted through theopenings at the ends 54 of the growth vessels 14 and 16. Then, culturemedium may be introduced into the growth vessels 14 and 16 through suchopenings at the ends, following which the openings may be closed by theplugs 18 and 20.

The culture medium in each of the growth vessels 14 and 16 is retainedin the reservoir formed by the flats 64 and having as its end walls thedam 62 and the rise 66, or if an excess of culture medium isinadvertently added, the neck 58 rather than the rise 66. This arrangement permits a gas, such as air or oxygen, to remain over the culturemedium which is in such reservoir.

Alternatively, the use of a cover slip may be eliminated, and cellcultures can be initiated on the floor of the flats 64.

The parabiotic chamber apparatus may be used to determine the transferof pathogenic conditions through the dialysis membrane, as byinoculating a tissue culture is growth Vessel 14 with a particularfiltrable virus and determining whether changes are effected in ahealthy tissue culture in growth vessel 16.

If desired, the dialysis membrane may be removed for specific researchprojects, and the flanges 60 of the growth vessels 14 and 16 may beoperatively engaged without the intermediate dialysis membrane 22.

In research problems Where the use of a gas above the liquid culturemedium is to be avoided, the embodiment shown in FIGURE 4 is used. Inthis embodiment, the holder 12 is identical to that employed in theembodiment of FIGURES 1, 2 and 3. The growth vessels 14a and 16aresemble the growth vessels 14 and 16 except that the darn 62 iseliminated, and an elbow 68 is provided intermediate the flat 64a andthe tapered end 54a. Thus, in this embodiment, the liquid culture mediummay be added to the growth vessel 14a so as to fill or substantiallyfill the same prior to the insertion of the plug 18a in the tapered end54a.

While the apparatus of the present invention has been described ashaving utility for tissue culture, it is to be understood that it isalso exceedingly useful in research in colloidal chemistry, and thelike, involving studies of unequal diifusion through membranes.

The present invention may be embodied in other specific forms withoutdeparting from the spirit or essential attributes thereof and,accordingly, reference should be made to the appended claims, ratherthan to the foregoing specification as indicating the scope of theinvention.

I claim:

1. A parabiotic chamber apparatus, comprising: a pair of vessels, eachof a tubular configuration open at both ends and having a flat floorextending along a central bottom portion between said open ends, saidfloor having a transverse width across said configuration and a longitudinal length extending between two constrictions of saidconfiguration arising from said floor to form boundaries between saidfloor and open ends, said width and length defining with saidconstrictions a culture growth area therein, means supporting saidvessels adjacent and aligned with one another for communicationtherebetween through adjacent open ends of the vessels; a dialysismembrane interposed between said adjacent ends; and plug members closingthe remote open ends of the vessels, said means comprising a horizontalbase and a holder means extending above said base for clamping saidadjacent ends together and supporting said vessels with said floor ofeach vessel at an elevated horizontal plane over said base.

2. A parabiotic chamber apparatus, comprising: a pair of vitreousvessels, each of an elongated tubular configuration open at both endsand having a flat floor extending along a central bottom portion betweensaid open ends, said floor having a transverse width substantially equalto a diameter through said tubular configuration and a longitudinallength extending between two constrictions of said configuration arisingfrom said floor to form boundaries between said floor and open ends,said width and length defining with said constrictions a culture growtharea therein; means supporting said vessels adjacent and aligned withone another for communication therebetween through adjacent open ends ofthe vessels; a dialysis membrane interposed between said adjacent ends;and plug members closing the remote open ends of the vessels, said meanscomprising a holder including a horizontal base and a vertical clampmember extending above said base, said member clamping said adjacentends together and supporting said vessels with said floor of each vesselat an elevated horizontal plane over said base.

3. A parabiotic chamber apparatus in accordance with claim 2 whereinsaid configuration includes an elbow portion directed upward to a remoteone of said open ends to thereby provide said vessels with said remoteends at an uppermost level.

4. A parabiotic chamber apparatus in accordance with claim 2 whereinsaid configuration includes an outward flared portion directed to anadjacent one of said open ends, thereby providing said vessels withadjacent ends of large cross-sectional area.

References Cited by the Examiner UNITED STATES PATENTS 1,331,732 2/1920Wait 2l0321 X 2,251,083 7/1941 Theorell 20430l X 2,276,986 3/1942 Kemp210-22 X 2,397,438 3/1946 Schmid 285423 X 3,055,504 9/1962 Schultz210321 X 3,069,340 12/1962 Mindick 2l0321 X 3,129,476 4/ 1964 Sindlinger285-423 X REUBEN FRIEDMAN, Primary Examiner.

THEODORE C. CRAVER, Examiner.

1. A PARABIOTIC CHAMBER APPARATUS, COMPRISING: A PAIR OF VESSELS, EACHOF A TUBULAR CONFIGURATION OPEN AT BOTH ENDS AND HAVING A FLAT FLOOREXTENDING ALONG A CENTRAL BOTTOM PORTION BETWEEN SAID OPEN ENDS, SAIDFLOOR HAVING A TRANSVERSE WIDTH ACROSS SAID CONFIGURATION AND ALONGITUDINAL LENGTH EXTENDING BETWEEN TWO CONSTRICTIONS OF SAIDCONFIGURATION ARISING FROM SAID FLOOR TO FORM BOUNDARIES BETWEEN SAIDFLOOR AND OPEN ENDS, SAID WIDTH AND LENGTH DEFINING WITH SAIDCONSTRICTIONS A CULTURE GROWTH AREA THEREIN, MEANS SUPPORTING SAIDVESSELS ADJACENT AND ALIGNED WITH ONE ANOTHER FOR COMMUNICATIONTHEREBETWEEN THROUGH ADJACENT OPEN ENDS OF THE VESSELS; A DIALYSISMEMBRANE INTERPOSED BETWEEN SAID ADJACENT ENDS; AND PLUG MEMBERS CLOSINGTHE REMOTE OPEN ENDS OF THE VESSELS, SAID MEANS COMPRISING A HORIZONTALBASE AND A HOLDER MEANS EXTENDING ABOVE SAID BASE FOR CLAMPING SAIDADJACENT ENDS TOGETHER AND SUPPORTING SAID VESSDLS WITH SAID FLOOR OFEACH VESSEL AT AN ELEVATED HORIZONTAL PLANE OVER SAID BASE.